广西植物2025，Vol.45Issue(9)：1679-1690,12.DOI:10.11931/guihaia.gxzw202403021
马缨杜鹃RdNAC72基因的克隆、亚细胞定位及表达分析
Cloning,subcellular localization,and expression analysis of the RdNAC72 gene in Rhododendron delavayi
摘要
Abstract
NAC transcription factors play important roles in plant growth,development,and stress responses.However,the molecular mechanism of the RdNAC72 gene in Rhododendron delavayi involved in response to stress remains unclear.To investigate the the roles of the RdNAC72 gene in high temperature stress response and its potential regulation mechenism,we first designed primers for cloning the full length coding sequence of the RdNAC72 gene using PCR technology.Subsequently,the gene's structure,function,and physicochemical properties were analyzed and predicted using bioinformatics method.The spatial and temporal expression characteristics of the RdNAC72 gene under heat stress and expression pattern under ABA stress were analyzed using quantitative real-time PCR(RT-qPCR).The results were as follows:(1)The RdNAC72 gene had a full length of 1 005 bp,encoding 334 amino acids with a relative molecular weight of 37.415 kDa.Subcellular localization analysis showed that the RdNAC72 protein was located in the nucleus.(2)Multiple sequence alignment and phylogenetic analysis indicated that the RdNAC72 was most closely related to the RwNAC72 in R.williamsianum.Additionally,cis-acting element analysis revealed that the gene contained elements associated with hormone response,light response,anaerobic response,low temperature response,and heat stress response.(3)High temperature stress could induce the expression of RdNAC72,exhibiting temporal and spatial expression specificity.After three days of high temperature stress treatment,the relative expression level of the RdNAC72 gene in leaves was significantly upregulated by 31.16-fold;while no significant change was observed in roots and stems.After six days of high temperature stress treatment,the relative expression levels of RdNAC72 roots,stems,and leaves were all significantly upregulated,with the highest observed in leaves(61.56-fold),followed by stems(50.14-fold),and roots(17.42-fold).Additionally,it was found that ABA could induce the expression of RdNAC72.(4)RT-qPCR analysis demonstrated a coordinated expression pattern between RdHSP17.2 and RdNAC72 with RdHSP17.2 containing multiple NAC recognition motifs(CATGTG)and core binding sequences(CACG)in its promoter region,indicating that RdNAC72 may be a downstream target gene of RdNAC72.Therefore,the RdNAC72,a transcription factor,localized in the nucleus,responds significantly to high temperature and ABA,potentially activating the RdHSP17.2 expression to confer heat resistance.This study enriches the biological functions of NAC transcription factors,and provides the reference for the future genetic and breeding of this plant.关键词
马缨杜鹃/RdNAC72/基因克隆/亚细胞定位/表达分析/高温胁迫Key words
Rhododendron delavayi/RdNAC72/gene cloning/subcellular localization/expression analysis/high temperature stress分类
农业科技引用本文复制引用
周平,王洪飞,韦云凤,胡晓玉,周玉梅,王孝敬..马缨杜鹃RdNAC72基因的克隆、亚细胞定位及表达分析[J].广西植物,2025,45(9):1679-1690,12.基金项目
国家自然科学基金(32260097) （32260097）
贵州省科技计划项目(黔科合中引地[2023]009). （黔科合中引地[2023]009）
